From: Methylation of the KEAP1 gene promoter region in human colorectal cancer
Methods | Primers | Sequence |
---|---|---|
BSP | Â | Forward: 5'-AAGAAAAGAAAAGAAAAGAAAATTTAG-3' |
 |  | Reverse: 5'-TTTAGTGAGGTAGATAATTTTTT-3' |
PCR conditions |  | Initial denaturation at 95°C (10 min) and 35 cycles at 95°C (3 s), 52°C (3 s), 72°C (5 s), and a final extension at 72°C (10 s) |
MSP | Methylation- | Forward: 5'-TAGATAATTTTTTTTAGATTTTGCGGTCG-3' |
 | Specific | Reverse: 5'-TCCTCGCGAAACTACGC-3' |
PCR condition |  | Initial denaturation at 95°C (10 min) and annealing temperature decrement of 0.5°C every cycle (from 70°C to 66.5°C) followed by 32 cycles of 66°C (3 s), 72°C (5 s), and a final extension at 72°C (10 s) |
MSP | Non-methylation | Forward: 5'-TAGATAATTTTTTTTAGATTTTGTGGTTG-3' |
 | -specific | Reverse: 5'-TCCTCACAAAACTACAC-3' |
PCR condition |  | Initial denaturation at 95°C (10 min) and annealing temperature decrement of 0.5°C every cycle (from 64°C to 60.5°C) followed by 32 cycles of 60°C(3 s), 72°C (5 s), and a final extension at 72°C (10 s) |